脱磷脂牛血红蛋白的制备

Preparation of Bovine Lipid-free Hemoglobin

研究了一种新的脱磷脂血红蛋白制备方法。在 2 %、5 %PEG40 0 0或 2 %、5 %PEG10 0 0 0作共溶剂的情况下 ,利用疏水相互作用色谱基本上完全除去了新鲜牛血红细胞裂解液中的磷脂类成分 ,其中以 5 %PEG40 0 0为共溶剂时 ,血红蛋白的回收率最高 ,达 85 .0 % ,在血红蛋白存在下所用的苯基琼脂糖 6B型疏水介质的吸附容量为 86 .6mg磷脂 mL介质。经过疏水作用色谱后 ,血红蛋白的P5 0是 3386 4Pa,Hill系数是 2 .5 4,较好地保存了血红蛋白的生物活性。

A new method for preparation of Hb solution free of stromal lipid was described. Almost all the lipid in fresh hemolysate of bovine red blood was removed with hydrophobic interaction chromatography (HIC) in the presence of 2%PEG4000, 5%PEG4000, 2%PEG10000 or 5%PEG10000. With the adding of 5%PEG4000, the 80% of recovery of Hb in HIC was obtained and the maximum lipid absorbed by hydrophobic medium, butyl agarose -6B was 86.6mg/mL. The activity (P50) of hemoglobin preparation was 3386.4Pa torrs, and the Hill number was 2.54, which were near to that of the native red blood cells. The mechanism of removing lipid by HIC and the function of PEG in the process were discussed.